Figure 3.

Novel ALK resistance mutations confer resistance PF02341066 and TAE684. A, polyclonal Ba/F3 cultures resistant to PF02341066 were treated with increasing concentrations of PF02341066. Viability was determined after 96 hours by measurements of cellular ATP content and expressed as a function of compound dose relative to dimethyl sulfoxide (DMSO)-treated controls. B, polyclonal Ba/F3 cultures resistant to PF02341066 were treated with increasing concentrations of TAE684. Viability was determined and depicted (as in A). C, Ba/F3 cells stably expressing EML4–ALK^wt or EML4–ALK^L1198P were treated with 1 µmol/L of PF02341066 or 500 nmol/L of TAE684. Cells were stained with trypan blue, and the fraction of viable cells was counted after 48 hours of treatment. D, whole-cell lysates of Ba/F3 stably expressing EML4–ALK^wt or EML4–ALK^L1198P were treated with different concentrations of ALK inhibitors. Levels of ALK phosphorylation were monitored by immunoblotting. Actin was used as loading control. wt, wild-type.